Vitamin D receptor gene polymorphisms: Analysis of ligand binding and hormone responsiveness in cultured skin fibroblasts

Recent reports have suggested that polymorphisms in the gene encoding the vitamin D receptor (VDR) determine a portion of the genetic contribution to bone mineral density (BMD). Individuals homozygous for the allele lacking the Bsm I restriction site in the intron between exons 8 and 9 (BB genotype) have been found to have lower BRID than individuals homozygous for the allele having the Bsm I site (bb genotype). Interestingly, this polymorphism has also been associated with prostate cancer risk, The observed changes in BMD and prostate cancer risk might be due to an alteration in the function or abundance of the VDR leading to differential responsiveness of target cells to the action of 1,25-dihydroxyvitamin D-3 [1,25(OH)(2)D-3]. To test this hypothesis, we cultured dermal fibroblasts from donors with BE, Bb, and bb genotypes and determined the level of VDR expression and the cellular responsiveness to 1,25(OH)(2)D-3 treatment, VDR abundance, affinity for [H-3]1,25(OH)(2)D-3, and VDR mRNA levels were not detectably different in BB cells compared to bb cells, Moreover, equal expression of both VDR gene alleles was detected by reverse transcriptase - polymerase chain reaction (RT-PCR) on mRNA from Bb fibroblasts. Fibroblast responsiveness to 1,25(OH)(2)D-3, assessed by induction of 24-hydroxylase mRNA, was similar between BE and bb cell types in dose-response experiments. Although there were individual variations in the parameters we measured, there were no detectable or consistent differences in mean values from our small sample of cultured dermal fibroblasts. In conclusion, we did not detect significant differences in VDR properties or cellular responsiveness to 1,25(OH)(2)D-3 that correlated with VDR genotype, Our findings suggest that these polymorphisms do not affect VDR function, but rather may be a marker for a nearby gene that is responsible for the genotype-associated variation in osteoporosis and prostate cancer risk. (C) 1998 Academic Press.